A method for isolating and culturing placental cells from failed early equine pregnancies.

Early pregnancy loss occurs in 6-10% of equine pregnancies making it the main cause of reproductive wastage. Despite this, reasons for the losses are known in only 16% of cases. Lack of viable conceptus material has inhibited investigations of many potential genetic and pathological causes. We present a method for isolating and culturing placental cells from failed early equine pregnancies. Trophoblast cells from 18/30 (60%) failed equine pregnancies of gestational ages 14-65 days were successfully cultured in three different media, with the greatest growth achieved for cells cultured in AmnioChrome™ Plus. Genomic DNA of a suitable quality for molecular assays was also isolated from 29/30 of these cases. This method will enable future investigations determining pathologies causing EPL.

Diseases in neonatal foals. Part 1: the 30 day incidence of disease and the effect of prophylactic antimicrobial drug treatment during the first three days post partum.

REASONS FOR PERFORMING STUDY: Neonatal diseases have been grouped and analysed but up-to-date statistically significant information about the incidence and prevalence of diseases in foals is limited. Since the 1950s it has been a common management practice to administer a 3 day course of antimicrobial drugs to neonatal foals. This was shown to significantly reduce the incidence of infections (Platt 1977). Since then management practices have improved and it is widely believed that prophylactic antimicrobial drugs are no longer necessary in foal rearing. OBJECTIVES: To determine the 30 day incidences or prevalences (depending on case definition) of various diseases and conditions in the neonatal foal and ascertain the influence of a prophylactic 3 day treatment on the frequency of infections. METHODS: The population consisted of Thoroughbred foals born on stud farms in the Newmarket (UK) area in 2005 (n = 1031). Depending on the stud farm's practice in the use of prophylactic antimicrobial drugs, 2 groups of newborn foals (treated and untreated) were identified and followed for 30 days. RESULTS: The 30 day incidences of infectious diseases under study were between 0.2% (osteomyelitis) and 5.85% (systemic disease with diarrhoea). The overall incidence for 'total infectious diseases' was 8.27%. The most commonly observed noninfectious condition was limb deformities (12.11% of all foals). There was no significant difference in the incidence of infectious diseases between the 2 groups. CONCLUSION: Infectious diseases are still an important problem in neonatal foals requiring further investigation as to which factors other than antimicrobial prophylaxis are relevant for disease prevention. POTENTIAL RELEVANCE: The results provide an up-to-date overview about the frequencies of various neonatal foal diseases. They do not support the traditional prophylactic use of antimicrobials to prevent infectious diseases in healthy newborn foals. However, it should be noted that this study was not a randomised controlled trial and therefore does not provide the strongest possible evidence for this conclusion.

Diseases in neonatal foals. Part 2: potential risk factors for a higher incidence of infectious diseases during the first 30 days post partum.

REASON FOR PERFORMING STUDY: The development of clinical illness in foals is usually predetermined by perinatal history, management or stressful environmental conditions. OBJECTIVES: To determine potential risk factors for an increased incidence of infectious diseases during the first 30 days post partum. METHODS: The population consisted of Thoroughbred foals born on stud farms in the Newmarket (UK) area in 2005 (n = 1031). They were followed for their first 30 days. Factors suspected to influence the incidence of infectious neonatal diseases were examined in a logistic regression approach for each of the 3 outcomes (total infectious diseases, systemic disease with diarrhoea and total infectious diseases excluding diarrhoea). All 28 factors were either foal or mare or stud farm related. RESULTS: Several significant risk factors for a higher disease incidence, such as birth complications, colostrum intake by stomach tube and leucocytosis 12-48 h post partum were identified. The factor 'boarding stud' seemed to be protective against disease. CONCLUSION: Some factors, such as the mare's time at stud before foaling, the mare's rotavirus vaccination schedule and fibrinogen-values that empirically had been linked to the outcome previously were not confirmed as relevant. This included the reported useful prophylactic treatment with antimicrobial drugs. POTENTIAL RELEVANCE: Factors to be considered when evaluating newborn foals include: stud management, the birth process, route of colostrum intake, white and red blood cells, and the date of birth. These may help to detect foals at risk to develop an infection so that targeted prophylactic measures can be initiated.

Physiological responses in nonheat acclimated horses performing treadmill exercise in cool (20 degrees C/40% RH), hot dry (30 degrees C/40% RH) and hot humid (30 degrees C/80% RH) conditions.

The aim of the present study was to determine the effect of different environmental conditions on physiological response to exercise. Four winter acclimatised, nonheat acclimated horses of different breeds were exercised at 20 degrees C/40% RH (CD), 30 degrees C/40% RH (HD) and 30 degrees C/80% RH (HH). The exercise test was designed to represent the structure and intensity of a One star Speed and Endurance test (competition exercise test [CET]). All 4 horses were able to complete the full CET (60 min + 30 min active recovery) in CD and HD, but only one horse completed the CET in HH. Two horses were stopped because of pronounced general fatigue and one because of a right atrial temperature (TRA) of 43 degrees C. Oxygen uptake on each phase was not different between CD and HD, but was higher during Phases B, C and D in HH. Mean peak TRA at the end of Phase D was 40.3 +/- 0.2, 41.6 +/- 0.4 and 42 +/- 0.3 degrees C for CD, HD and HH, respectively. Corresponding, mean peak rectal temperatures (TREC) following Phase D were 39.5 +/- 0.1, 40.6 +/- 0.1 and 41.5 +/- 0.1 degree C for CD, HD and HH, respectively. Mean time to peak TREC was 9.3 +/- 1.1 (CD), 7.3 +/- 1.8 (HD) and 10.8 +/- 2.3 (HH) min and was not significantly different between conditions (P > 0.05). Heat dissipation amounted to 83 +/- 1, 73 +/- 2 and 70 +/- 1% of heat production in CD, HD and HH, respectively. Weight loss was significantly correlated with both body surface area (CD r = 0.85; HD r = 0.87; HH r = 0.81) and bodyweight (CD r = 0.97; HD r = 0.93; HH r = 0.94). The greatest weight loss recorded was 4.6% bodyweight in one horse in HD. The mean increase in exercise intensity over the whole CET (in terms of VO2) of HD and HH and HH compared with CD was 5 +/- 3 and 14 +/- 3% higher, respectively. The exercise induced hyperthermia and the reduced capacity for heat dissipation produced partial compensatory responses in minute ventilation (VE), particularly during Phase C, when the horses were trotting. In HD, the increase in VE was achieved mainly through an increase in frequency, whilst in HH it was achieved through an increase in tidal volume (VT). The horses demonstrated a high degree of tolerance to environmental heat load, suggesting a high thermoregulatory capacity. However, for unacclimatised animals exercising in severely hot and humid conditions, performance may be limited.

Physiological, metabolic and biochemical responses of horses competing in the speed and endurance phase of a CCI*****3-day-event.

The present study was undertaken to investigate physiological, metabolic, haematological and biochemical changes in horses competing in the Speed and Endurance test of a Concours Complet International (CCI)*****3-day-event held under FEI rules. A total of 28 horses competing in the Burghley Horse Trials Speed and Endurance test were selected to be monitored: 11 horses in 1993 and 17 horses in 1994. Of the 28 horses selected, 17 completed the Speed and Endurance test and went on to complete the showjumping test. Mean +/- s.d. shade temperature and relative humidity, black globe temperature and wind speed were 13 +/- 1 and 20 +/- 2 degrees C, 54 +/- 3 and 55 +/- 10%, 17 +/- 2 and 29 +/- 4 degrees C and 2.7 +/- 0.7 and 1.2 +/- 0.3 m/s, for 1993 and 1994, respectively. Mean heart rate during Phases A, B and D was not significantly different between years, but mean heart rate during Phase C and X was significantly higher in 1994. Mean (+/- s.d.) heart rate on Phase B and D for all horses in both 1993 and 1994 was 198 +/- 8 and 188 +/- 11 beats/min, respectively. Mean heart rate during Phase D showed a poor correlation with mean speed (r = 0.412). Total mean (+/- s.d.) weight loss from the start of Phase A to the end of Phase D was 15.5 +/- 6.1 kg in 1993 and 16.5 +/- 5 kg in 1994 and did not differ significantly between years. Following 14-18 h completion of Phase D, mean bodyweight was not significantly different from that at the start of Phase A in either year. Mean rectal temperature at the end of Phase D was 41 +/- 0.6 degrees C and 41.1 +/- 0.6 degrees C in 1993 and 1994, respectively (P > 0.05). Both the lowest (39.7 degrees C) and highest (41.8 degrees C) rectal temperatures were recorded at the end of Phase D in 1994. Plasma lactate concentrations at the end of Phase D were 8.5-38.5 mmol/l. The highest lactate concentration also coincided with the highest plasma glucose concentration (11.4 mmol/l) as well as the joint fastest time in either year, although overall lactate showed only weak correlations with mean speed on Phase D (r = 0.12, 1993; r = 0.58, 1994). While the Speed and Endurance test at CCI*****level run in a temperate climate presents a considerable challenge to the fitness and ability of the horses competing, the metabolic and physiological changes are not extreme. The majority of horses that finish the test appear to undergo a rapid and considerable degree of recovery and are able to present sound at the final inspection, take part in the showjumping test and complete the competition.

Clinical observations made in nonheat acclimated horses performing treadmill exercise in cool (20 degrees C/40%RH), hot, dry (30 degrees C/40%RH) or hot, humid (30 degrees C/80%RH) conditions.

Four horses (H, J, N and M) undertook a treadmill competition exercise test (CET), designed to simulate the physiological and metabolic stresses of the Speed and Endurance phase of a 3-day-event, under 3 different environmental conditions: 20 degrees C/40% relative humidity (RH) (cool, dry [CD]: 2 sessions); 30 degrees C/40%RH (hot, dry [HD]) and 30 degrees C/80%RH (hot, humid [HH]) (Marlin et al. 1995). A number of subjective clinical observations were made at designated time points throughout the exercise test and initial recovery period including buccal mucous membrane colouration, capillary refill time, neck and point of shoulder skin pinch recovery time, grade of abdominal sounds; anal sphincter tone as well as the presence or absence of fatigue and ataxia. The aim was to investigate their value in predicting performance in the final canter phase of the CET equivalent to the cross-country or Phase D of a field competition. In addition, the use of a more objective assessment, the cardiac recovery index (CRI), was investigated together with the heart rate, rectal temperature and respiratory frequency at the end of Phase C and at the 8 min point of the 10 Minute Box (8'X). The CRI was calculated according to the formula CRI = P2-P1 where P2 = the heart rate in beats/min at the 8 min point of the '10 Minute Box' (Phase X) of the CET. P1 = the heart rate (beats/min) at the 7 min point just before the horse was made to trot over a distance of 80 m at a speed of 3.7 m/s (at a 3 degrees incline) before returning to a walk. The study suggested that the subjective tests carried out at the 'End-C' and/or '8'X' time points were not useful in predicting subsequent performance in the final canter phase (Phase D) and neither were heart rate, rectal temperature or respiratory frequency. However, the only horse (Horse H) to complete the full CET under HH conditions was the only animal to show a decrease in respiratory frequency between the End-C and 8'X time points. All others showed an obvious increase. Under HH conditions, Horse H also had the lowest CRI. For 3 of the horses the highest CRI value was found under the HH conditions, for the fourth horse an equally high CRI value was found with one of the CD sessions. However, under the HH conditions, both P1 and P2 values were > 100. The study suggested that it could be beneficial if a suitably modified CRI test, as well as a procedure to monitor the change in respiratory frequency during the 10 Minute Box, were evaluated further on the treadmill and in the field with respect to their potential usefulness as additional aids to the assessment of a horse's suitability to proceed to Phase D.